Expression and characterization of human tissue kallikrein variants

Protein Expression and Purification
H ChanJ M Clark

Abstract

Human tissue kallikrein is a serine protease implicated in the pathology of various inflammatory disorders. As one of the two principal enzymes that generate proinflammatory kinin peptides in vivo, tissue kallikrein represents an attractive target for therapeutic intervention in diseases such as asthma, pancreatitis, and rheumatoid arthritis. Three distinct human tissue kallikrein variants, differing in one or two amino acid substitutions, are predicted to exist based on genomic or cDNA nucleotide sequences derived from different tissues. The effects of these substitutions on the biochemical properties of tissue kallikrein are unknown but could, in principle, confer tissue-specific functions on the enzyme or affect the clinical utility of specific kallikrein inhibitors. All three variants, as well as a deglycosylated derivative, were expressed in high yield as recombinant proteins in Pichia pastoris. The recombinant kallikrein variants and natural urinary kallikrein all hydrolyzed synthetic peptides with similar specificity and efficiency and released kallidin from kininogen at comparable rates. Similarly, no significant differences were observed in the interactions between kallikrein variants and protein inhibitors such as SBT...Continue Reading

Citations

Jan 21, 2006·American Journal of Respiratory Cell and Molecular Biology·S Marina Casalino-MatsudaRosanna M Forteza
Jul 29, 2006·American Journal of Respiratory Cell and Molecular Biology·Rosanna FortezaAnthony J Day
Nov 2, 2018·Supportive Care in Cancer : Official Journal of the Multinational Association of Supportive Care in Cancer·Barbara Bonaparte Ribas de AlcântaraAuro Del Giglio
Apr 22, 2020·PloS One·Gwendolyn VliegenIngrid De Meester
May 6, 1998·Protein Science : a Publication of the Protein Society·B A KatzE B Springman
Feb 28, 2004·The Journal of Biological Chemistry·Susana M Casalino-MatsudaRosanna M Forteza
Jan 21, 2000·FEMS Microbiology Reviews·J L Cereghino, J M Cregg
Mar 27, 1999·Protein Expression and Purification·H ChanJ M Clark
Jun 19, 2001·Biochimica Et Biophysica Acta·M ZaniT Moreau
Mar 26, 2002·Journal of the American Society of Nephrology : JASN·Rola SlimFrançois Alhenc-Gelas

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